OBTAINING CLONE-SPECIFIC PRIMER AND PROBE FOR THE IMMUNOGLOBULIN HEAVY-CHAIN GENE FROM PARAFFIN-EMBEDDED TISSUE OF B-CELL LYMPHOMA - TECHNICAL CONSIDERATIONS

The complementarity determining region (CDR) III of the immunoglobulin heavy-chain (IgH) gene is a tumor-specific marker for B-cell malignan cies that has been widely exploited for the monitoring of minimal resi dual disease in B-precursor acute lymphocytic leukemia. There are a nu mber of technical problems in applying the same technology for B-cell non-Hodgkin's lymphoma (B-NHL). Several procedures have been useful in overcoming these unique problems encountered in obtaining the tumor-s pecific sequence of the IgH-CDRIII in B-NHL, including the use of dena turing gradient gel electrophoresis or micromanipulation of tissue sec tions in separating the tumor-specific CDRIII products from those of c ontaminating normal B-lymphocytes, Minor modifications of a commercial kit greatly improve the purity of the polymerase chain reaction (PCR) products for sequencing. Modifications of the 5'-ends of the V-H and I-H primers, coupled with the cycle sequencing technique, make it poss ible to obtain unambiguous sequences on direct sequencing of short PCR products. Computer informatics and programs that facilitate the desig n of tumor-specific primers and probes from CDRIII sequences are descr ibed.